Probing Activated Sludge with Fluorescently Labeled rRNA Targeted Oligonucleotides

1994 ◽  
Vol 29 (7) ◽  
pp. 15-23 ◽  
Author(s):  
M. Wagner ◽  
R. Amann ◽  
H. Lemmer ◽  
W. Manz ◽  
K. H. Schleifer
Keyword(s):  
Activated Sludge ◽  
Ribosomal Rna ◽  
Phosphate Removal ◽  
Municipal Sewage ◽  
Correct Identification ◽  
Rich Medium ◽  
Phylogenetic Groups ◽  
In Situ Investigation ◽  
Nutrient Rich Medium

Activated sludge samples from municipal sewage treatment plants were characterized using 16S and 23S ribosomal RNA targeted oligonucleotide probes specific for defined phylogenetic groups of bacteria. Comparison of in situ community structures as determined by molecular biological methods with the composition of the heterotrophic saprophyte flora isolated on nutrient rich medium revealed large discrepancies. These are caused by the selectivity of media and culture conditions. The most significant effect of cultivation on nutrient rich medium is an underestimation of bacteria belonging to the beta-subclass of Proteobacteria and an overestimation of bacteria belonging to the gamma-subclass of Proteobacteria. Therefore, culture dependent enumerations of the gamma-subclass bacteria of the genus Acinetobacter in plants with enhanced biological phosphate removal (EBPR) resulted in significant overestimations. In situ identification by fluorescent oligonucleotide probing revealed that Acinetobacter numbers were below 8% of the active bacterial cells in the examined EBPR-plants. In situ hybridization techniques also bear the potential for the early and correct identification of filamentous bacteria as indicators for sludge bulking and foaming. A 16S ribosomal RNA targeted oligonucleotide probe specific for Sphaerotilus spec, was developed and successfully applied for in situ investigation of this filamentous bacterium.

Application of oligonucleotide probes for the detection of Thiothrix spp. in activated sludge plants treating paper and board mill wastes

2002 ◽  
Vol 46 (1-2) ◽  
pp. 559-564 ◽  
Author(s):  
S.B. Kim ◽  
M. Goodfellow ◽  
J. Kelly ◽  
G.S. Saddler ◽  
A.C. Ward
Keyword(s):  
Activated Sludge ◽  
Bacterial Population ◽  
Oligonucleotide Probe ◽  
In Situ Hybridisation ◽  
Filamentous Bacteria ◽  
Phylogenetic Groups ◽  
Investigation Period ◽  
Mixed Populations ◽  
Conventional Microscopy

Filamentous bacteria belonging to the genus Thiothrix were detected in activated sludge samples using the fluorescent in situ hybridisation (FISH) technique. A 16S rRNA-targeted oligonucleotide probe was developed for the detection of members of the T. fructosivorans group, and the performance of probe TNI for the detection of Thiothrix nivea group was enhanced by using an unlabeled competitor. A set of 5 probes covering all phylogenetic groups of Thiothrix were used to examine samples taken from selected activated sludge plants treating paper and board mill wastes. Members of the T. eikelboomii group formed the predominant filamentous bacterial population in plants experiencing poor sludge settleability, whereas members of the T. nivea group were commonly found but not dominantly in the remaining plants. Members of the T. fructosivorans group were not detected at any significant level in any of the samples. The distribution of the main Thiothrix types remained unchanged throughout the investigation period. It was evident that mixed populations of Thiothrix spp. were present in all activated sludge samples investigated, the observed differences were in the relative abundance of the various groups. These findings were supported by the results obtained using conventional microscopy.

scholarly journals Filamentous Bacterium Eikelboom Type 0092 in Activated Sludge Plants in Australia Is a Member of the Phylum Chloroflexi

2009 ◽  
Vol 75 (8) ◽  
pp. 2446-2452 ◽  
Author(s):  
Lachlan Speirs ◽  
Tadashi Nittami ◽  
Simon McIlroy ◽  
Sarah Schroeder ◽  
Robert J. Seviour
Keyword(s):  
In Situ Hybridization ◽  
Activated Sludge ◽  
Molecular Data ◽  
Phosphate Removal ◽  
Filamentous Bacterium ◽  
Eastern Australia ◽  
Intracellular Aggregates ◽  
Rrna Sequences ◽  
16S Rrna Sequences

ABSTRACT Molecular data show that the filamentous bacterium Eikelboom type 0092, frequently seen in Australian activated sludge plants, is a member of the phylum Chloroflexi. Fluorescence in situ hybridization (FISH) probes designed against cloned 16S rRNA sequences from a full-scale enhanced biological phosphate removal-activated sludge plant community, where this was a dominant filament morphotype, suggest that it can exist as two variants, differing in their trichome diameter. When applied to samples from several treatment plants in eastern Australia, each FISH probe targeted only the type 0092 filament morphotype against which it was designed. The patterns of FISH signals generated with both were consistent with the ribosomes not being evenly distributed but arranged as intracellular aggregates. The FISH survey data showed that these two variants appeared together in most but not all of the plants examined. None stained positively for intracellular presence of either poly-β-hydroxyalkanoates or polyphosphate.

16S rRNA in situ probing for the determination of the family level community structure implicated in enhanced biological nutrient removal

2001 ◽  
Vol 43 (1) ◽  
pp. 91-98 ◽  
Author(s):  
D. D. Mudaly ◽  
B. W. Atkinson ◽  
F. Bux
Keyword(s):  
Activated Sludge ◽  
Nutrient Removal ◽  
Phosphate Removal ◽  
Microbial Community Analysis ◽  
Biological Nutrient Removal ◽  
Total Bacterial Count ◽  
Family Level ◽  
Acinetobacter Spp ◽  
Alpha Proteobacteria

Knowledge of a discrete physiological group capable of excess biological phosphate removal (EBPR) remains unclear. Consequently, microbial community analysis of an enhanced continuous laboratory-scale activated sludge process displaying a strong EBPR mechanism was conducted. Unit design was configured upon the three-stage Phoredox process and characterization of the activated sludge bacterial community was carried out using fluorescent in situ hybridization (FISH) techniques. Fixed activated sludge samples were hybridized with fluorescently labeled oligonucleotide probes targeting the following bacterial phylogenetic divisions: a kingdom level probe specific for all bacteria (EUB338); family level probes specific for the alpha, beta and gamma subclasses of the class Proteobacteria; Gram positive bacteria with a high (G+C) DNA content (GPBHGC) or Actinobacteria; the Cytophaga-Flavobacterium (CF) subclass within the Cytophaga-Flavobacterium-Bacteriodes division; and genus level probes specific for Pseudomonas spp., Aeromonas spp., and Acinetobacter spp. Bacterial predominance between the anaerobic, anoxic and aerobic zones of the EBPR sludge were comparable and appeared as follows; beta (22%), alpha (19%), gamma (17%), GPBHGC (11%) and CF (8%). The incidence of Acinetobacter spp. appeared to be generally low with counts amounting to <9% of the total bacterial count. A population shift in the alpha Proteobacteria subclass was evident between the non-nutrient removal seed sludge and the EBPR sludge, implicating this group in EBPR. The overall results indicate that the beta and alpha Proteobacteria can be metabolically functional in EBPR processes and reiterate the functional misconception of Acinetobacter spp. in EBPR systems.

In situ investigation of the primary recrystallization of alpha titanium in HVEM

1978 ◽  
Vol 36 (1) ◽  
pp. 634-635
Author(s):  
S. Naka ◽  
R. Penelle ◽  
R. Valle
Keyword(s):  
Dimensional Analysis ◽  
Texture Evolution ◽  
Cold Work ◽  
Three Dimensional ◽  
Primary Recrystallization ◽  
Thin Foils ◽  
In Situ Investigation ◽  
Grain Growth Model ◽  
Alpha Titanium

The in situ experimentation technique in HVEM seems to be particularly suitable to clarify the processes involved in recrystallization. The material under investigation was unidirectionally cold-rolled titanium of commercial purity. The problem was approached in two different ways. The three-dimensional analysis of textures was used to describe the texture evolution during the primary recrystallization. Observations of bulk-annealed specimens or thin foils annealed in the microscope were also made in order to provide information concerning the mechanisms involved in the formation of new grains. In contrast to the already published work on titanium, this investigation takes into consideration different values of the cold-work ratio, the temperature and the annealing time.Two different models are commonly used to explain the recrystallization textures i.e. the selective grain growth model (Beck) or the oriented nucleation model (Burgers). The three-dimensional analysis of both the rolling and recrystallization textures was performed to identify the mechanismsl involved in the recrystallization of titanium.

In-situ Investigation of Bainite Formation with fast X-Ray Diffraction (iXRD)

2017 ◽  
Vol 72 (6) ◽  
pp. 355-364
Author(s):  
A. Kopp ◽  
T. Bernthaler ◽  
D. Schmid ◽  
G. Ketzer-Raichle ◽  
G. Schneider
Keyword(s):  
X Ray Diffraction ◽  
X Ray ◽  
In Situ Investigation

Behaviour of Alkylphenol Polyethoxylate Surfactants and of Nitrilotriacetate in Sewage Treatment

1987 ◽  
Vol 19 (3-4) ◽  
pp. 449-460 ◽  
Author(s):  
W. Giger ◽  
M. Ahel ◽  
M. Koch ◽  
H. U. Laubscher ◽  
C. Schaffner ◽  
...  
Keyword(s):  
Activated Sludge ◽  
Sewage Treatment ◽  
Operating Conditions ◽  
Municipal Sewage ◽  
Sludge Treatment ◽  
Loading Rates ◽  
Treatment Conditions ◽  
Physico Chemical ◽  
Treated Sewage ◽  
Removal Efficiencies

Effluents and sludges from several municipal sewage treatment plants in Switzerland were analyzed for nonylphenol polyethoxylates (NPnEO, n=3-20), nonylphenol mono- and diethoxylate (NPlEO, NP2EO), corresponding nonylphenoxy carboxylic acids (NP1EC, NP2EC) and nonylphenol (NP). These chemicals derive from nonionic surfactants of the NPnEO-type, and specific analitical techniques were used to study their behaviour during mechanical-biological sewage and subsequent sludge treatment. The parent NPnEO-surfactants, with concentrations in raw and mechanically treated sewage from 400-2200 mg/m3, were relatively efficiently removed by the activated sludge treatment. The abundances of the different metabolites varied depending on treatment conditions. The refractory nature of NPl/2EO, NP and NPl/2EC was recognized. Both biotransformations and physico-chemical processes determine the behaviour and fate of nonylphenolic substances in sewage treatment. Nitrilotriacetate (NTA) was found in primary effluents at concentrations between 430 and 1390 mg/m3. The various treatment plants showed different removal efficiencies for NTA depending on the operating conditions. Activated sludge treatment with low sludge loading rates and nitrifying conditions removed NTA with efficiencies between 95 and 99%. High sludge loading caused a decrease in NTA removal efficiencies from 70% to 39%.

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